Structure-function studies of the bHLH phosphorylation domain of TWIST1 in prostate cancer cells

Rajendra P Gajula; Sivarajan T Chettiar; Russell D Williams; Katriana Nugent; Yoshinori Kato; Hailun Wang; Reem Malek; Kekoa Taparra; Jessica Cades; Anvesh Annadanam; A-Rum Yoon; Elana Fertig; Beth A Firulli; Lucia Mazzacurati; Timothy F Burns; Anthony B Firulli; Steven S An; Phuoc T Tran

doi:10.1016/j.neo.2014.10.009

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Structure-function studies of the bHLH phosphorylation domain of TWIST1 in prostate cancer cells

Journal article

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Structure-function studies of the bHLH phosphorylation domain of TWIST1 in prostate cancer cells

Rajendra P Gajula, Sivarajan T Chettiar, Russell D Williams, Katriana Nugent, Yoshinori Kato, Hailun Wang, Reem Malek, Kekoa Taparra, Jessica Cades, Anvesh Annadanam, …

Neoplasia (New York, N.Y.), Vol.17(1), pp.16-31

01/2015

DOI: https://doi.org/10.1016/j.neo.2014.10.009

PMID: 25622896

Abstract

Prostatic Neoplasms - metabolism

TOR Serine-Threonine Kinases - metabolism

Humans

Protein Multimerization

Transcriptome

Male

Gene Expression Profiling

Phosphatidylinositol 3-Kinases - metabolism

Structure-Activity Relationship

Epithelial-Mesenchymal Transition - genetics

Cell Movement - genetics

Neoplasm Metastasis

Heterografts

Prostatic Neoplasms - genetics

Basic Helix-Loop-Helix Transcription Factors - metabolism

Protein Interaction Domains and Motifs

Nuclear Proteins - genetics

Proto-Oncogene Proteins c-akt - metabolism

Twist-Related Protein 1 - chemistry

Disease Models, Animal

Prostatic Neoplasms - pathology

Basic Helix-Loop-Helix Transcription Factors - genetics

Nuclear Proteins - metabolism

Nuclear Proteins - chemistry

Amino Acid Motifs

Cell Movement - drug effects

Animals

Signal Transduction - drug effects

Twist-Related Protein 1 - genetics

Cell Line, Tumor

Protein Kinase Inhibitors - pharmacology

Twist-Related Protein 1 - metabolism

Basic Helix-Loop-Helix Transcription Factors - chemistry

Cluster Analysis

Phosphorylation

Mutation

The TWIST1 gene has diverse roles in development and pathologic diseases such as cancer. TWIST1 is a dimeric basic helix-loop-helix (bHLH) transcription factor existing as TWIST1-TWIST1 or TWIST1-E12/47. TWIST1 partner choice and DNA binding can be influenced during development by phosphorylation of Thr125 and Ser127 of the Thr-Gln-Ser (TQS) motif within the bHLH of TWIST1. The significance of these TWIST1 phosphorylation sites for metastasis is unknown. We created stable isogenic prostate cancer cell lines overexpressing TWIST1 wild-type, phospho-mutants, and tethered versions. We assessed these isogenic lines using assays that mimic stages of cancer metastasis. In vitro assays suggested the phospho-mimetic Twist1-DQD mutation could confer cellular properties associated with pro-metastatic behavior. The hypo-phosphorylation mimic Twist1-AQA mutation displayed reduced pro-metastatic activity compared to wild-type TWIST1 in vitro, suggesting that phosphorylation of the TWIST1 TQS motif was necessary for pro-metastatic functions. In vivo analysis demonstrates that the Twist1-AQA mutation exhibits reduced capacity to contribute to metastasis, whereas the expression of the Twist1-DQD mutation exhibits proficient metastatic potential. Tethered TWIST1-E12 heterodimers phenocopied the Twist1-DQD mutation for many in vitro assays, suggesting that TWIST1 phosphorylation may result in heterodimerization in prostate cancer cells. Lastly, the dual phosphatidylinositide 3-kinase (PI3K)-mammalian target of rapamycin (mTOR) inhibitor BEZ235 strongly attenuated TWIST1-induced migration that was dependent on the TQS motif. TWIST1 TQS phosphorylation state determines the intensity of TWIST1-induced pro-metastatic ability in prostate cancer cells, which may be partly explained mechanistically by TWIST1 dimeric partner choice.

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Title: Structure-function studies of the bHLH phosphorylation domain of TWIST1 in prostate cancer cells
Creators: Rajendra P Gajula - Department of Radiation Oncology and Molecular Radiation Sciences, Sidney Kimmel Comprehensive Cancer Center, Johns Hopkins University School of Medicine, Baltimore, MD, USA
Sivarajan T Chettiar - Department of Radiation Oncology and Molecular Radiation Sciences, Sidney Kimmel Comprehensive Cancer Center, Johns Hopkins University School of Medicine, Baltimore, MD, USA
Russell D Williams - Department of Radiation Oncology and Molecular Radiation Sciences, Sidney Kimmel Comprehensive Cancer Center, Johns Hopkins University School of Medicine, Baltimore, MD, USA
Katriana Nugent - Department of Radiation Oncology and Molecular Radiation Sciences, Sidney Kimmel Comprehensive Cancer Center, Johns Hopkins University School of Medicine, Baltimore, MD, USA
Yoshinori Kato - The Russell H. Morgan Department of Radiology and Radiological Science, Division of Cancer Imaging Research, Johns Hopkins University School of Medicine, Baltimore, MD, USA; Department of Oncology, Sidney Kimmel Comprehensive Cancer Center, Johns Hopkins University School of Medicine, Baltimore, MD, USA; In Vivo Cellular and Molecular Imaging Center, Johns Hopkins University School of Medicine, Baltimore, MD, USA
Hailun Wang - Department of Radiation Oncology and Molecular Radiation Sciences, Sidney Kimmel Comprehensive Cancer Center, Johns Hopkins University School of Medicine, Baltimore, MD, USA
Reem Malek - Department of Radiation Oncology and Molecular Radiation Sciences, Sidney Kimmel Comprehensive Cancer Center, Johns Hopkins University School of Medicine, Baltimore, MD, USA
Kekoa Taparra - Department of Radiation Oncology and Molecular Radiation Sciences, Sidney Kimmel Comprehensive Cancer Center, Johns Hopkins University School of Medicine, Baltimore, MD, USA; Cellular and Molecular Medicine Program, Johns Hopkins University School of Medicine, Baltimore, MD, USA
Jessica Cades - Department of Radiation Oncology and Molecular Radiation Sciences, Sidney Kimmel Comprehensive Cancer Center, Johns Hopkins University School of Medicine, Baltimore, MD, USA
Anvesh Annadanam - Department of Radiation Oncology and Molecular Radiation Sciences, Sidney Kimmel Comprehensive Cancer Center, Johns Hopkins University School of Medicine, Baltimore, MD, USA
A-Rum Yoon - Department of Environmental Health Sciences, Johns Hopkins University Bloomberg School of Public Health, Baltimore, MD, USA
Elana Fertig - Department of Oncology, Division of Biostatistics and Bioinformatics, Sidney Kimmel Comprehensive Cancer Center, Johns Hopkins University School of Medicine, Baltimore, MD, USA
Beth A Firulli - Department of Pediatrics, Riley Heart Research Center, Indiana University School of Medicine, Indianapolis, IN, USA
Lucia Mazzacurati - Department of Medicine, Division of Hematology-Oncology, Hillman Cancer Center, University of Pittsburgh School of Medicine, Pittsburgh, PA, USA
Timothy F Burns - Department of Medicine, Division of Hematology-Oncology, Hillman Cancer Center, University of Pittsburgh School of Medicine, Pittsburgh, PA, USA
Anthony B Firulli - Department of Pediatrics, Riley Heart Research Center, Indiana University School of Medicine, Indianapolis, IN, USA
Steven S An - In Vivo Cellular and Molecular Imaging Center, Johns Hopkins University School of Medicine, Baltimore, MD, USA; Department of Environmental Health Sciences, Johns Hopkins University Bloomberg School of Public Health, Baltimore, MD, USA; Physical Sciences in Oncology Center, Johns Hopkins University, Baltimore, MD, USA; Department of Chemical and Biomolecular Engineering, Johns Hopkins University, Baltimore, MD, USA
Phuoc T Tran - Department of Radiation Oncology and Molecular Radiation Sciences, Sidney Kimmel Comprehensive Cancer Center, Johns Hopkins University School of Medicine, Baltimore, MD, USA; Department of Oncology, Sidney Kimmel Comprehensive Cancer Center, Johns Hopkins University School of Medicine, Baltimore, MD, USA; In Vivo Cellular and Molecular Imaging Center, Johns Hopkins University School of Medicine, Baltimore, MD, USA; Cellular and Molecular Medicine Program, Johns Hopkins University School of Medicine, Baltimore, MD, USA; Department of Urology, Johns Hopkins University School of Medicine, Baltimore, MD, USA. Electronic address: tranp@jhmi.edu
Publication Details: Neoplasia (New York, N.Y.), Vol.17(1), pp.16-31
Date published: 01/2015
Grant note: T32 CA121937 / NCI NIH HHS P30 CA006973 / NCI NIH HHS R01 HL122123 / NHLBI NIH HHS R01 HL120920 / NHLBI NIH HHS P50CA103175 / NCI NIH HHS U54CA141868 / NCI NIH HHS R01 HL107361 / NHLBI NIH HHS R01 AR061392 / NIAMS NIH HHS HL107361 / NHLBI NIH HHS R01CA166348 / NCI NIH HHS R43 CA141868 / NCI NIH HHS T32CA121937 / NCI NIH HHS 1R0AR061392 / NIAMS NIH HHS P50 CA103175 / NCI NIH HHS T32 GM008752 / NIGMS NIH HHS T32 CA009071 / NCI NIH HHS R01 CA166348 / NCI NIH HHS U54 CA143868 / NCI NIH HHS
Academic Unit: Clinical and Translational Research
Language: English
Resource Type: Journal article
Identifiers: 991031654594904646